生物物理学中的单分子技术：方法和应用进展综述,Reports on Progress in Physics

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生物物理学中的单分子技术：方法和应用进展综述
Reports on Progress in Physics ( IF 19.0 ) Pub Date : 2017-12-19 , DOI: 10.1088/1361-6633/aa8a02
Helen Miller ₁ , Zhaokun Zhou , Jack Shepherd , Adam J M Wollman , Mark C Leake

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单分子生物物理学改变了我们对生物学以及生命物理学的理解。比简单的软物质更奇特的是，生物物质远离热平衡，覆盖从单个分子的纳米级到细胞、组织和生物体中高达几个数量级的多个长度。生物分子通常以潜在的不稳定性为特征：存在多个亚稳态自由能态，仅由热能标度 k BT 的几个倍数的水平分隔，其中 k B 是玻尔兹曼常数和 T 绝对温度，这意味着复杂的相互转化动力学活性生物物质的相对炎热潮湿的环境。单分子生物物理学技术的一个关键优势是它们能够探测分子群中自由能状态的异质性，对于传统的整体平均方法来说，一般来说太具有挑战性了。实验和计算技术的并行发展促进了多路复用相关技术的诞生，以解决以前难以解决的生物学问题。在实验上，检测器灵敏度和速度的提高以及光源、探针和微流体的稳定性和效率的提高推动了进展。我们讨论了这些近期实验的动机和要求，包括基础数学。这些方法大致分为检测分子的工具和操纵分子的工具。对于前者，我们讨论了超分辨率显微镜的进展，对于解决生命科学中许多长期存在的问题具有变革性，对于后者，我们将进展包括在“ 力谱技术，机械地扰动分子。我们还考虑了单分子计算物理学的计算机进展，以及如何将模拟和实验结合在一起以提供更完整的理解。现在越来越多地使用组合技术，包括相关原子力显微镜和荧光成像，以探索更接近天然生理行为的问题。我们确定了这些技术的权衡、限制和应用，并讨论了令人兴奋的新方向。包括相关原子力显微镜和荧光成像，以探索更接近天然生理行为的问题。我们确定了这些技术的权衡、限制和应用，并讨论了令人兴奋的新方向。包括相关原子力显微镜和荧光成像，以探索更接近天然生理行为的问题。我们确定了这些技术的权衡、限制和应用，并讨论了令人兴奋的新方向。

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Single-molecule techniques in biophysics: a review of the progress in methods and applications

Single-molecule biophysics has transformed our understanding of biology, but also of the physics of life. More exotic than simple soft matter, biomatter lives far from thermal equilibrium, covering multiple lengths from the nanoscale of single molecules to up to several orders of magnitude higher in cells, tissues and organisms. Biomolecules are often characterized by underlying instability: multiple metastable free energy states exist, separated by levels of just a few multiples of the thermal energy scale k B T, where k B is the Boltzmann constant and T absolute temperature, implying complex inter-conversion kinetics in the relatively hot, wet environment of active biological matter. A key benefit of single-molecule biophysics techniques is their ability to probe heterogeneity of free energy states across a molecular population, too challenging in general for conventional ensemble average approaches. Parallel developments in experimental and computational techniques have catalysed the birth of multiplexed, correlative techniques to tackle previously intractable biological questions. Experimentally, progress has been driven by improvements in sensitivity and speed of detectors, and the stability and efficiency of light sources, probes and microfluidics. We discuss the motivation and requirements for these recent experiments, including the underpinning mathematics. These methods are broadly divided into tools which detect molecules and those which manipulate them. For the former we discuss the progress of super-resolution microscopy, transformative for addressing many longstanding questions in the life sciences, and for the latter we include progress in 'force spectroscopy' techniques that mechanically perturb molecules. We also consider in silico progress of single-molecule computational physics, and how simulation and experimentation may be drawn together to give a more complete understanding. Increasingly, combinatorial techniques are now used, including correlative atomic force microscopy and fluorescence imaging, to probe questions closer to native physiological behaviour. We identify the trade-offs, limitations and applications of these techniques, and discuss exciting new directions.

更新日期：2017-12-19

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